Pasteurella multocida is a Gram‐negative bacterium that causes hemorrhagic septicemia (HS) in buffaloes and cattle. The disease causes serious problems in Indonesian livestock and is classified as a serious transmissible animal disease. Previous research has determined the diversity of P. multocida using a serotyping method based on the antigenic properties of capsule polysaccharides. An alternative method for analysis utilizes sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and random amplified polymorphic DNA (RAPD). This study aimed to characterize and determine P. multocida diversity in several regions of Indonesia based on phenotypic character, protein profile, and the band pattern of RAPD results. Bacterial identification was performed using traditional biochemical techniques and API® 20NE systems and then confirmed molecularly using polymerase chain reaction (PCR). The freeze‐thawing technique was performed to obtain the bacterial protein extract, and DNA extraction was executed using DNAzol. The extracted protein and RAPD product were then electrophoresed on 12% polyacrylamide gel and 1.5% agarose gel, respectively. The results indicate that the molecular weight range of the protein bands is 12–209 kDa, and the band pattern of the RAPD results ranged from 307–3,100 bp. Based on phenotypical analysis, P. multocida from South Sulawesi Province exhibited a variety of growth characteristics in MacConkey agar media. Using the hierarchical clustering analysis of the band patterns of RAPD and the whole‐cell protein profiles, four and five clusters were formed, respectively. These results indicate molecular diversity among P. multocida from several regions of Indonesia.

Juwita S, Indrawati A, Damajanti R, Safika, Mayasari NLPI. 2021. Multiple antibiotic resistance and virulence factors of Staphylococcus aureus strains isolated from dairy farms in South Sulawesi, Indonesia. Biodiversitas 23: 1015-1022. Antimicrobial resistance (AMR) is an important issue affecting human and animal health worldwide. This study aimed to investigate antibiotic resistance and determine the virulence factors of S. aureus isolated from the dairy farms in South Sulawesi, Indonesia. Thirty-one isolates of S. aureus were tested for sensitivity to 9 types of antibiotics using the Kirby-Bauer disk diffusion method. The analysis of antibiotic resistance and virulence genes in S. aureus isolates was performed by the conventional PCR method. The results showed that S. aureus isolates from human samples were resistant to penicillin G (PEN) (86%), ampicillin (AMP) (86%), oxacillin (OXA) (14%), cefoxitin (FOX) (14%), tetracycline (TET) (43%) and ciprofloxacin (CIP) (14%). Staphylococcus aureus isolates from the animal samples were resistant to penicillin G (PEN) (50%), ampicillin (AMP) (50%), tetracycline (TET) (15%), and erythromycin (5%). Meanwhile, S. aureus isolates from dangke were resistant to penicillin G (PEN) and ampicillin (AMP) (50% each). Antimicrobial resistance genes for blaTEM (83%), mecA (17%), and tetA (100%) were detected in S. aureus isolates from human samples, whereas those for blaTEM (90%) and tetA (100%) were detected in isolates from animal samples. Meanwhile, the genes for blaTEM (100%) were detected in isolates from dangke. A total of 19 S. aureus isolates harbored the virulence gene for fnbA (26%), clfA (58%), hla (58%), and tst (21%). The use of antibiotics in humans and animals needs to be implemented properly in local communities to prevent the spread of antibiotic resistance. The presence of the tst gene in raw milk is essential for consumer protection against the risk of toxic shock syndrome.

Background and Aim: Orangutans are an “umbrella species” for conserving tropical forests in Sumatra and Kalimantan. There are remarkable changes between the gut microbiomes of wild and captive Sumatran orangutans. This study aimed to profile gut microbiota of wild and captive Sumatran orangutans. Materials and Methods: Nine fecal samples collected from wild orangutans and nine fecal samples collected from captive orangutans were divided into three replicates. Each replicate randomly combined three pieces and were analyzed on the Illumina platform. A bioinformatics study of 16S rRNA according to Qiime2 (Version 2021.4) and microbiome profiling analysis was conducted. Results: The relative abundance of different microbial taxa varied significantly between wild and captive Sumatran orangutans. Among the operational taxonomic units, various proportions of Firmicutes, Proteobacteria, Bacteroidetes, Euryarchaeota, Acidobacteria, Actinobacteria and Verrucomicrobia predominated. Solobacterium was found only in 19% of captive orangutans. Methanobrevibacter was identified to be prevalent among wild orangutans (16%). Analysis of the core microbiome from the combined wild and captive data revealed seven species as cores. According to linear discriminant analysis effect size, Micrococcus luteus, Bacteroidescaccae, Lachnospiraceae bacterium, Ruthenibacterium lactatiformans, Haemophilus haemolyticus, and Chishuiella spp. were microbiome biomarkers in captive orangutans, whereas Roseburia inulinivorans, Collinsella aerofaciens, Oscillibacter spp., and Eubacterium hallii were microbiome biomarkers in wild orangutans. Conclusion: There were differences in the microbiome biomarkers of wild and captive Sumatran orangutans. This study is important for understanding the role of gut bacteria in the health of Sumatran orangutans. Keywords: captive Sumatran orangutans, core microbiome, gut microbiota, Pongo abelii, wild Sumatran orangutans.

Aim: This study aimed to isolate and identify lactic acid bacteria (LAB) in wild Sumatran orangutans to provide more information about LAB diversity derived from Sumatran orangutan feces. Materials and Methods: Fecal sampling from three female orangutans, around 35 years old, was carried out in the wild forest areas at the research station of Suaq Belimbing Gunung Leuser National Park located in the South Aceh district. Orangutan fecal samples were taken in the morning when the orangutans first defecated. The orangutans were above the tree, which is approximately 12-15 m from the ground where feces were found. Results: Fermentation testing using the API 50 CHL Kit showed that OUL4 isolates were identified as Lactococcus lactis ssp. lactis with an identity value of 73.5%. Homology analysis demonstrated that the OUL4 isolates have 93% similarity to Weissella cibaria, and phylogenetic trees constructed using Mega 7.0 also showed that OUL4 isolates are related to W. cibaria. Conclusion: These results show that there is a difference in identification between biochemical testing with API kits and molecular analyses on LAB isolates from wild Sumatran orangutans. Based on 16S rRNA gene homology, the OUL4 LAB isolates from wild Sumatran orangutans have 93% homology to W. cibaria.

Background and Aim: Staphylococcus aureus is a bacterium that causes several infectious diseases, including mastitis, endocarditis, and osteomyelitis, and poses a threat to human and animal health. This study aims to phenotypically and genetically identify S. aureus from the isolates collected from humans, animals, environment, and Dangke products in the dairy farms of South Sulawesi Province, Indonesia, as well as to establish a genetic relationship among the isolated S. aureus strains. Materials and Methods: The total number of samples was 142, comprising 30 humans (skin swab), 58 animals (raw milk), 14 dairy products (Dangke), and 40 environmental samples (water). S. aureus was phenotypically identified using the culture method, followed by Gram staining, catalase test, and coagulase test. Simultaneously, genotypic identification of S. aureus was performed using the conventional polymerase chain reaction and sequencing methods. Sequencing data were analyzed using the MEGA X software by comparing BLAST National Center for Biotechnology Information databases. Results: The phenotypic methods revealed that 56/142 (39.4%) animal, human, and Dangke samples grew on culture, and 56/56 (100%) were Gram stain positive, 56/56 (100%) catalase-positive, and 23/56 (41.1%) coagulase positive. The genotypic method revealed that 32/56 (57.1%) samples amplified the nuc gene. The phylogenetic analysis of 12 isolates revealed that they are all closely related and do not belong to distinct clades. Conclusion: It indicates that S. aureus isolates from animals (S30) are probably the same strain as human isolates (H2, H3, H4, and H5). The findings of this study can be used as information regarding the importance of preventing and controlling diseases caused by S. aureus using a health approach involving the human, animal, and environmental sectors. This study was limited to the sequencing analysis of the nuc gene.

This study was aimed to identify possible critical points of Escherichia coli (E. coli ) O157:H7, a pathogenic agent, contamination in aceh cattle breeding centre. For this purpose, samples were collected from cattle faeces, hand of workers (animal keepers), and water sources in the farm using cross-sectional approach. A number of 85 samples of cattle faecal swab were collected randomly from the animals in the breeding centre. The samples of swab of hand of all workers (15 persons) were collected before and after work. Then, the water sources from 11 cattle house locations in the breeding centre were collected. The water sources were divided into three different locations, namely the water containers, taps, and water puddle on the floors. At each source a number of 11 samples were collected. Isolation of E. coli was conducted on Eosin Methylene Blue Agar (EMBA), followed by identification on Sorbitol MacConkey Agar (SMAC). Then Molecular subtyping of E. coli O157:H7 genes was conducted using multiplex-PCR analysis. Data were analysed descriptively. The results of this study showed that 72 samples (85 %) among 85 samples were positive for E. coli and the rest of samples were positive for other types of bacteria. Sample isolation from swabs of hand was found 3 positive E. coli before work and 1 positive E. coli after work from 15 workers. The most potential water sources for E. coli contamination were the water in taps, and water puddle on the floor of cattle houses. Then, two of samples of E. coli isolated from rectal swab were confirmed as E. coli O157:H7 using PCR test, based on the presence of stx2 gene. In conclusion, the risk of presence of E. coli as zoonotic agents of E. coli O157:H7 in aceh cattle as well as from the farm workers and surrounding area are high. An appropriate control strategy is needed to apply in the aceh cattle farm to prevent from E. coli O157:H7 outbreak in the future.